Anti hladr antibodies

Lymphocytes isolated from peripheral blood, LN, jejunum, and rectum were quantified for their activation (CD38+, CD69+, and HLADR+), proliferation (Ki67+), and naïve/memory phenotype (CD28+ and CD95+). Cells were first stained with live/dead stain (Thermo Fisher Scientific, Eugene, OR, USA). After washing, cells were further stained with cocktail of monoclonal antibodies (mAbs) including anti-CD3, anti-CD4, anti-CD8, anti-CD28, anti-CD95, anti-CD38, anti-CD69, and anti-HLADR antibodies (BD Biosciences, San Jose, CA, USA; Table S2) as reported earlier. To detect proliferating Ki67+ cells, intracellular staining protocol was performed using BD Fix/Perm solutions as described earlier [26 (link)]. Cells were fixed with 1x BD stabilizing and fixative buffer. At least 20,000 events were collected by gating on lymphocytes from each sample, and the data were analyzed using FlowJo software, version 10.7.0. (Becton, Dickinson & Company, Franklin Lakes, NJ, USA).

Flow cytometric analysis was performed on a FACS-Calibur analytical flow cytometer using CellQuest-Pro software (BD Biosciences). DC maturation markers were detected by staining with PE-labelled anti-CD83 and APC-labelled anti-HLA-DR antibodies (BD Biosciences).